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CHAPTER 6:

CHAPTER 6:

CHAPTER 6: GENERAL CONCLUSIONGENERAL CONCLUSIONGENERAL CONCLUSIONGENERAL CONCLUSION

The work described in this thesis investigated new procedures that could be

used to synthesise melanin in a reproducible fashion and to test the

effectiveness of sunscreens containing melanin or other sunscreen agents and

characterise skin colour by diffuse reflectance.

Synthesis of melanins Synthesis of melanins Synthesis of melanins Synthesis of melanins

Despite a large number of studies incorporating synthetic melanins, relatively

few studies have examined melanin synthesis techniques as a means of

synthesising melanin reproducibly.

With the method of melanin synthesis outlined in this study melanins were

synthesised in batches. As melanins were synthesised in batches synthesis

variables, such as reaction temperature, condenser water temperature, room

temperature and ambient light, between the batches were minimised.

The synthesis procedure outlined was simple and inexpensive and the

variables were easily monitored and controlled. The method also allowed for

the synthesis of multiple batches of melanin simultaneously in a reproducible

fashion.

Characterisation of melanins Characterisation of melanins Characterisation of melanins Characterisation of melanins

Conversion of the melanin pyrograms to “pseudo-mass spectra” allowed the

objective comparison of melanin batches using software commercially

available for the comparison of mass spectra.

From the results of this study, melanins produced from different precursors

could be differentiated the basis of statistically significant differences in their

pyrograms. Conversely, statistically significant differences between

pyrograms of different batches of melanins produced under the same

conditions from the same precursors could not be demonstrated. The

analytical technique in this study could be used in combination with other

analytical techniques to determine if the melanins synthesised by the methods

outlined in this thesis were reproducible or as the basis of future studies

examining differences between melanins synthesised from the same precursor

but under different conditions.

Characterisation of skin colour by di Characterisation of skin colour by di Characterisation of skin colour by di

Characterisation of skin colour by diffuse reflectanceffuse reflectanceffuse reflectanceffuse reflectance

This section of work was undertaken to provide a method for assessing the

ability of sunscreen formulations to adhere to the skin. As a melanin

containing sunscreen is likely to be brown, the ability of a formulation to

adhere to skin could be monitored by objectively measuring changes in the

skin colour as the product was washed or worn off the skin.

A spectrophotometer was modified to measure diffuse reflectance outside of

the sample compartment. The modifications to the spectrophotometer

spectrophotometers, relatively inexpensive, reliable, and which could

accurately quantify changes in the colour of human skin

This device developed compares favourably with similar devices described in

the literature in terms of reproducibility and has the advantages of shorter

scan times and a higher resolution. It was simple to construct and at about

A$600 was approximately one tenth the cost of similar commercial devices.

In vitro In vitro In vitro

In vitro evaluati evaluati evaluati evaluations of Sun Protective Factors of sunscreen agentons of Sun Protective Factors of sunscreen agentons of Sun Protective Factors of sunscreen agentons of Sun Protective Factors of sunscreen agent

In vivo methods are the accepted method for the determination of SPF values of sunscreens. However, in vivo methods for determination of the SPF values of sunscreens formulations are time consuming and expensive. In vitro

methods have the potential to give rapid results at a fraction of the cost of

in vivo methods. Unfortunately, in vitro methods often correlate poorly with

in vivo results and are generally unable to determine high SPF values accurately due to limitations in the ability of in vitro methods to measure a wide range of light levels.

The in vitro study presented here used human stratum corneum and Transpore surgical tape as substrates, which were easily obtained. The

in vitro method was fast, simple and capable of measuring high SPF

sunscreens (>70). Although more data in the high SPF range (>30) would be

required to confirm results in this range. The value of this technique was

in vivo testing. The corrected value was subsequently confirmed by independent in vivo testing.

Few naturally occurring pigments command more widespread interest than

melanins [22]. As the development of novel pharmaceutical products

containing melanin increases, methods for the synthesis and characterisation

of melanin containing products will become increasingly important.

Furthermore, there is currently a high consumer demand for sunscreen

products that offer better protection from sun damage, due in part to public

education campaigns and the increased understanding of the mechanisms

and long term effects of sun exposure. The development of fast, inexpensive

and reliable techniques for ascertaining the efficacy of sunscreens will hasten

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